ABSTRACT
Three selective enrichment broths - selenite cystine (SC), Muller-Kauffmann tetrathionate (MKT) and Rappaport-Vassiliadis (RV) - were compared, for Salmonella Typhimurium isolation from rectal swabs of a calf experimentally infected. The bacteriological procedure involved pre-enrichment in Hajna-GN broth (only for the samples inoculated in RV broth), selective enrichment (SC, MKT and RV broths), culture in modified brilliant green agar (BGA), presumptive biochemistry tests (using triple-sugar-iron agar and lysine-agar) and slide agglutination test with poli-O and poli-H Salmonella antisera. SC and MKT broths were more efficient in the isolation of Salmonella Typhimurium (12 positive samples), whereas RV broth had a lower efficiency in the microbiological isolation (ten positive samples).(AU)
Subject(s)
Animals , Cattle , Salmonella Infections, Animal , Culture Media/analysis , Salmonella typhimurium/isolation & purification , Agar , Agglutination TestsABSTRACT
Three selective enrichment broths - selenite cystine (SC), Muller-Kauffmann tetrathionate (MKT) and Rappaport-Vassiliadis (RV) - were compared, for Salmonella Typhimurium isolation from rectal swabs of a calf experimentally infected. The bacteriological procedure involved pre-enrichment in Hajna-GN broth (only for the samples inoculated in RV broth), selective enrichment (SC, MKT and RV broths), culture in modified brilliant green agar (BGA), presumptive biochemistry tests (using triple-sugar-iron agar and lysine-agar) and slide agglutination test with poli-O and poli-H Salmonella antisera. SC and MKT broths were more efficient in the isolation of Salmonella Typhimurium (12 positive samples), whereas RV broth had a lower efficiency in the microbiological isolation (ten positive samples).
ABSTRACT
ABSTRACT This work was carried out to assess the preenrichment (PE) and enrichment (DE) steps for isolating Salmonella serotypes Enteritidis (SE) and Typhimurium (STM) from chicken feces kept at 4° C for 24 and 96h. The samples were artificially contaminated and kept in 1% peptone water at 4° C for 24 or 96h. After that, part of them was incubated at 37° C/24h and part was inoculated into enrichment broth, selenite broth plus novobiocin (SN) and tetrathionate broth plus novobiocin (TN) incubated at 37°C/24h. The PE culture was inoculated in SN, TN and Rapapport-Vassiliadis novobiocin (RVN), also incubated at 37° C/24h. The enrichment broth was plated on brilliant green agar (BGA), MacConkey agar (MCA), Hektoen agar (HEA), Salmonella-Shigella agar (SSA), xylose-lysine desoxicholate agar (XLDA) and xylose-lysine tergitol 4 (XLT4), which were incubated at 37° C/24h. Salmonella-like colonies were submitted to TSI agar and LIA agar, and incubated at 37° C/24h, as well as to slide agglutination tested with poly O and poly H Salmonella antiserum. When the samples were stored for 24h there was no difference between PE and DE (p > 0.05). However after 96h the PE was superior to DE (p 0.05). For enrichment, better results were seen with RVN broth (p 0.05). The XLD yielded
RESUMO Este trabalho foi desenvolvido para avaliar comparativamente o isolamento de Salmonella sorotipos Enteritidis (SE) e Typhimurium (STM) a partir do enriquecimento direto (ED) ou processamento com pré-enriquecimento (PE) de amostras de fezes de aves adultas, armazenadas em água peptonada tamponada a 1% (APT) por 24 ou 96h a 4º C. Utilizou-se os caldos de enriquecimento Rapapport-Vassiliadis novobiocina (RVN), tetrationato-novobiocina (TN) e selenitonovobiocina (SN) e os meios para plaqueamento ágar verde brilhante (VB), ágar de MacConkey (MC), ágar de Hektoen (HE), ágar Salmonella-Shigella (SS), ágar xilose lisina desoxicolato (XLD) e ágar xilose lisina tergitol 4 (XLT4). O procedimento bacteriológico incluiu as etapas de pré-enriquecimento, enriquecimento em caldo seletivo, plaqueamento, testes bioquímicos presuntivos e confirmação sorológica com utilização de soros polivalentes anti-antígenos somáticos e anti-antígenos flagelares de Salmonella. Não houve diferença estatisticamente significativa (p > 0,05) para as amostras armazenadas por 24h submetidas tanto ao PE quanto ao ED. Entretanto, em armazenagem por 96h o número de isolamentos nas amostras submetidas ao PE foi estatisticamente superior às submetidas ao ED (p 0,05). Quanto aos caldos enriquecedores, não houve diferença estatística de número de isolamentos (p > 0,05) entre os caldos SN e TN, mas o caldo RVN mostrou-se estatisticamente superior aos demais (p 0,05). Para os meios de plaqueamento, o XLD destacou-se por promover maior número de recuperações, embora sem significado estatístico (p > 0,05) para as amostras estocadas por 24h. Entre os dois sorotipos de Salmonella (SE e STM) não houve diferença estatística no número de recuperações (p > 0,05).
ABSTRACT
We evaluated the effectiveness of Selenite Cystine (SC), Tetrathionate Brilliant Green (TBG) and Rappaport Vassiliadis (RV) broths for Salmonella isolation. We also tested three classic plating media, Salmonella-Shigella Agar (SS), Brilliant Green Agar (BGA), Xylose Lysine Desoxycholate Agar (XLD) and two chromogenic agars, Rambach (RA) and CHROMagar Salmonella (CAS). Among 100 poultry carcasses, 29 were positive for Salmonella using all plating media combined. RV broth (69%) and TT broth (58.6%) were more effective than SC broth (24.1%). The chromogenic media gave better results than the classic ones with less false-positive colonies. The most effective isolation medium was CHROMagar, where Salmonella was identified in 23 (79.3%) of the 29 positive samples, followed by Rambach (48%). Positivity for Salmonella using classic media was 13.8% for BGA, 27.6% for SS and 34.5% for XLD.
A eficiência dos caldos selenito cistina (SC), tetrationato verde brilhante (TBG) e Rappaport Vassiliadis (RV) foi avaliada quanto ao isolamento de Salmonella. Também foram testados três meios clássicos de isolamento, ágar Salmonella-Shigella (SS), ágar Verde Brilhante (VB) e ágar xilose lisina desoxicolato (XLD) e dois meios cromogênicos, Rambach (RA) e CHROMagar (CAS). Entre 100 carcaças de frango examinadas, 29 foram positivas para Salmonella usando todos os meios combinados. Os caldos RV (69%) e TT (58,6%) foram mais eficientes que o SC (24,1%). Os meios cromogênicos mostraram melhores resultados do que os clássicos na detecção de Salmonella e apresentaram uma quantidade menor de colônias falso-positivas. O meio cromogênico mais eficiente foi o CAS, que detectou Salmonella em 23 das 29 amostras positivas (79,3%), seguido pelo RA (48%). Entre os meios clássicos, a detecção foi de 13,8% para VB, 27,6% para SS e 34,5% para XLD.
ABSTRACT
A two-phase study was conducted to compare the efficacy of several enrichment selective-broth steps associated to different plating media for recovery of Salmonella sp. from finishing swine feces. In a first phase, Rappaport-Vassiliadis broth (RV) incubated at 42ºC, Tetrathionate Müller-Kauffmann broth at 37ºC (TMK37) and 42ºC (TMK42), and Selenite Cystine broth (SC) at 37ºC, in combination with three selective plating media Rambach agar (RA), Xylose-Lysine-Tergitol 4 agar (XLT4), and Brilliant-Green Phenol-Red Lactose Sucrose agar (VB) were compared for recovery of Salmonella from artificially contaminated swine feces. In a second phase, RV, TMK37, and TMK42, associated with XLT4 and VB , were tested with naturally contaminated swine feces. In this study RV, TMK42 and TMK37 were superior to SC for isolating Salmonella sp. from artificially contaminated feces. TMK42 and RV were more productive than TMK37 for recovery of Salmonella from naturally contaminated feces samples. Selectivity and indication capability of the plating media were remarkably affected by the selective enrichment step effectiveness. The TMK42/XLT4 association was the most sensitive and RV/XLT4 the most specific. The use of VB agar is also recommended to increase the likelihood of isolating atypical H2S-late producing/ non-producing Salmonella. In this study RV and TMK42 were the most efficient selective enrichment for recovery of Salmonella sp. from swine feces.
Através de um estudo em duas fases comparou-se a eficiência de etapas de enriquecimento seletivo, associadas a diferentes meios seletivos, na recuperação de Salmonella sp. de fezes de suínos de terminação. Em uma primeira fase, as amostras foram contaminadas artificialmente e os caldos Rappaport-Vassiliadis (RV) incubado a 42ºC, Tetrationato Müller-Kauffmann a 37ºC (TMK37) e 42ºC (TMK42), e Selenito Cistina (SC) a 37ºC foram testados, em associação com meios sólidos seletivos: ágar Rambach (RA), ágar Xilose-Lisina-Tergitol 4 (XLT4), e ágar Verde-brilhante Vermelho-neutro Lactose Sacarose (VB). Na segunda fase os caldos RV, TMK37, and TMK42, semeados nos meios XLT4 and VB, foram testados com amostras naturalmente contaminadas. No isolamento de Salmonella sp. em amostras artificialmente contaminadas o RV, TMK42 e TMK37 foram superiores ao SC. Na segunda fase o TMK42 e RV foram mais eficientes que o TMK37. O desempenho destas etapas de enriquecimento seletivo influenciou diretamente a capacidade seletiva e indicadora dos meios sólidos seletivos utilizados. No presente estudo, a associação TMK42/XLT4 demonstrou ser a mais sensível, e a RV/XLT4 a mais específica. O uso do meio VB também é recomendado para aumentar a probabilidade do isolamento de colônias atípicas de Salmonella - produtoras tardias ou não produtoras de H2S. No presente estudo, RV e TMK42 foram as etapas de enriquecimento seletivo mais eficientes para o isolamento de Salmonella de fezes de suínos.